magistrsko delo
Saša Kobal (Author), Simon Horvat (Mentor), Simon Horvat (Thesis defence commission member), Primož Klinc (Thesis defence commission member), Mojca Narat (Thesis defence commission member), Gregor Majdič (Thesis defence commission member), Primož Klinc (Co-mentor)

Abstract

Liniji FLI (angl. Fat, F) in FHI (angl. Lean, L) sta liniji, ki sta nastali z divergentno selekcijo v 60 generacijah na lastnosti povezani z maso telesnega maščevja. Na koncu selekcijskega poskusa je imela linija F visok (~24) odstotek telesnega maščevja, L-linija pa ~4 %. Zaradi ekstremov v deležu telesnega maščevja imata liniji slabše z reprodukcijo povezane lastnosti, kar predstavlja en vidik ogroženosti ohranjanja teh linij samo z vzrejo. Drug vidik ogroženosti je dejstvo, da je reja vezana samo na Oddelek za zootehniko Biotehniške fakultete Univerze v Ljubljani in obstaja možnost, da v primeru bolezni in drugih nesreč ta dva genetska vira izgubimo. Zato je bil glavni namen naloge razviti protokole zamrzovanja za morebitno potrebo po ponovni oživitvi linij. V prvem delu smo želeli optimizirati protokol superovulacije samic z optimizacijo časovnega intervala med aplikacijo hormonov hCG in PMSG. V drugem delu smo preučevali uspešnost pridobivanja dvoceličnih zarodkov in vitro pri linijah L, F ter kontrolnih linijah CD-1 in B6D2F1, ki smo jih po gojenju in vitro zamrzovali z metodo vitrifikacije. Zarodke smo odmrznili in jih gojili v in vitro pogojih do faze blastociste. Pri liniji F se je razvoj ustavil pri štiriceličnem stadiju. Pri liniji L pa se je pri 3 % dvoceličnih zarodkov razvoj nadaljeval do stadija blastociste. Presaditev odmrznjenih dvoceličnih zarodkov linije L je uspela z do 19 % odstavljenih mladičev po kotitvi, za linijo F pa ni uspela. V drugem sklopu magistrske naloge smo želeli vzpostaviti protokol za zamrzovanje semenčic linije F in L, ampak nam kljub modifikacijam ni uspelo razviti učinkovitega protokola. Pri zamrzovanju zarodkov debele linije F in pri postopkih zamrzovanja semenčic linij nismo bili uspešni. Pri selekcijski liniji L pa smo uspeli zamrzniti zarodke in pridobiti žive mladiče po odmrznitvi, in vitro gojenju in presaditi zarodkov.

Keywords

biotehnologija;zamrzovanje zarodkov;optimizacija postopkov zamrzovanja;presaditev zarodkov;mišja linija L;

Data

Language: Slovenian
Year of publishing:
Typology: 2.09 - Master's Thesis
Organization: UL BF - Biotechnical Faculty
Publisher: [S. Kobal]
UDC: 606:61:57.086.13 (043.2)
COBISS: 13492483 Link will open in a new window
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Downloads: 159
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Other data

Secondary language: English
Secondary title: Optimization of methods for embryo cryopreservation to conserve and manage the endangered mouse selection lines fli (''fat'') and fhi (''lean'')
Secondary abstract: The FLI (Fat, F) and FHI (Lean, L) lines were developed by divergent selection over 60 generations on properties related to body fat. At the end of the selection experiment, the F-line had a high (~ 24) percentage of body fat and the L-line had only ~ 4%. Due to extreme in body fat these lines have poor reproduction and are hence endangered if maintained only by breeding. Another threat lies in the fact that breeding is only done at the Department of Animal Science, Biotechnical Faculty, University of Ljubljana in Slovenia and hence there is a possibility that in the case of diseases and disasters, these two genetic resources may be lost. Therefore, the main purpose of this Master’s thesis was to develop cryopreservation protocols for the potential need for line rederivation. In the first part, we wanted to optimize the protocol of female superovulation by optimizing the time interval between hCG and PMSG hormone application. In the second part, we tested protocols for in vitro production of two-cell embryos in line L, F, and control lines CD-1 and B6D2F1. 2-cell stage embryos were in vitro cultivated and frozen by vitrification. The embryos were thawed and cultured under in vitro conditions until the blastocyst stage. In line F, development stopped at the four-cell stage. For line L, however, in 3% of the two-cell embryos, development continued to the blastocyst stage. We were able to obtain up to 19% of weaned pups of L lineage, but these procedures failed for the F- line. In the second part of the Master's thesis we wanted to establish a protocol for cryopreservation of sperm in lines F and L, but despite modifications we were not able to develop an effective protocol. In summary, we were unsuccessful in cryopreservation of F lineage embryos and in sperm freezing procedures. In the selection line L, however, we succeeded in freezing the embryos and obtaining live pups after thawing, in vitro cultivation and embryo transfer.
Secondary keywords: biotechnology;cryopreservation of embryos;optimisation of cryopreservation procedures;, embryotransfer;mouse line L;
Type (COBISS): Master's thesis/paper
Study programme: 0
Embargo end date (OpenAIRE): 1970-01-01
Thesis comment: Univ. v Ljubljani, Biotehniška fak., Študij biotehnologije
Pages: IX, 43 f.
ID: 11609699